Furin

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Furin
Furin
Ribbon model by Furin, according to PDB  1P8J
other names
  • Paired Basic Amino Acid Cleaving Enzyme (PACE)

Existing structure data : 1P8J , 4OMC , 4OMD , 4RYD , 4Z2A

Properties of human protein
Mass / length primary structure 794 aa , 86,678 Da
Secondary to quaternary structure Homotetramer
Cofactor Ca 2+
Isoforms 1 (+ 4 potential [computer-aided])
Identifier
Gene name FURINE
External IDs
Enzyme classification
EC, category 3.4.21.75 ,  hydrolase
MEROPS S08.071
Response type hydrolysis
Substrate Precursor protein
Products mature protein + peptide
Occurrence
Homology family HOG000192536
Parent taxon Bilateria
Orthologue
human House mouse
Entrez 5045 18550
Ensemble ENSG00000140564 ENSMUSG00000030530
UniProt P09958 P23188
Refseq (mRNA) NM_002569 NM_001081454
Refseq (protein) NP_001276752 NP_001074923
Gene locus Chr 15: 90.87 - 90.88 Mb Chr 7: 80.39 - 80.41 Mb
PubMed search 5045 18550

Furin is an endoprotease of the proprotein convertase family (PC) that catalyzes the proteolytic maturation of precursor proteins in the eukaryotic protein secretion pathway . It is a ubiquitously expressed Type I transmembrane protein that is found in all vertebrates and many invertebrates . The substrates include growth factors , receptors , extracellular matrix proteins and also other protease systems that control certain diseases. In addition to activating pathogens , it also plays an essential role in embryogenesis and homeostasis .

The diseases in which furin is involved include anthrax , avian flu , cancer , Alzheimer's disease and Ebola fever , among others .

structure

The large extracellular regions of furin share an overall homology with the same regions of other members of the proprotein convertase family that belong to the superfamily of subtilisin- like serine proteases . The greatest sequence similarities are in the subtilisin-like catalytic domain . The aspartate , histidine and serine residues , which together form the catalytic triad , are highly conserved and the catalytic domains of other proprotein convertases are 54–70% identical to the sequence of furin. In addition to a signal peptide , furin and other proprotein convertases have so-called prodomains, where the interface of the signal peptide at the N terminus and several conserved basic amino acid residues which contain the autoproteolytic interface at the C terminus are located . This essential pro-domain plays an important role in PC protein folding , activation, transport and regulation. Furin and other PCs also have a P-domain , which is necessary for enzyme activity, adjustment to the pH value and for the recruitment of calcium as a cofactor . The P-domain is absent in bacterial PCs. The cytoplasmic domain of furin controls its localization and protein sorting (the process by which proteins are sorted based on their destination) in the trans -Golgi network .

properties

Furin has a broad pH optimum. 50% of its enzymatic activity is between pH 5 and 8, depending on which substrate is cleaved. Like other members of the subtilisin superfamily, furin is strictly dependent on calcium and requires approximately 1 mM calcium for complete function. Furin also has two calcium binding pockets, one with medium affinity and the other with high affinity . Furin also has a weak bond with potassium ; a potassium concentration of 20 mM increases the furin activity by increasing the deacylation rate (reverse reaction of the acylation ), which is important in the catalytic cycle of furin.

The consensus interface at which furin cleaves is located behind the arginine residue at the C terminus in the sequence - Arg - X - Lys / Arg - Arg - | - (where X is any amino acid, the vertical line with the quarter-quarters is the interface , the slash denotes an " or link " and the double- quarter line denotes a peptide bond ) and was determined biochemically with two furin substrates, one with the protective antigen ( PA for short , a subunit of anthrax toxin ) and the other with the hemagglutinin of influenza virus A. (HA). The arginine residues at the P1 position (amino acid residue which is N -terminal to the interface) and P4 position (four amino acid residues in the N -terminal direction from the interface) are essential, whereas the basic amino acid at the P2 position is not is essential, but can strongly influence the efficiency of the enzymatic conversion. Therefore, the sequence - Arg - X - X - Arg - | - represents the minimum requirement for a cutting sequence for furin, with non-preferred residues at the P4 position being compensated by preferred amino acid residues at the P2 and P6 positions. Because of this, the sequence - Lys / Arg - X - X - X - Lys / Arg - Arg - | - also represents a cutting sequence for furin in exceptional cases.

The two most widely used furin inhibitors are the stoichiometric peptidyl inhibitor decanoyl - Arg - Val - Lys - Arg - chloromethyl ketone and the α 1 -antitrypsin Portland (α 1 -PDX), a biotechnologically produced variant of the α 1 -antitrypsin . Decanoyl – Arg – Val – Lys – Ar – chloromethyl ketone inhibits all PC with a low nanomolar inhibition constant ( K i ), whereby the alkylating properties of the reactive group limit the application possibilities of the reagent. α 1 -PDX is generated by mutation at a reactive point on a loop, so that the minimum requirement for a cutting sequence for furin is met (- Arg - Ile - Pro - Arg - | -). In addition, α 1 -PDX is highly selective for furin in vitro ( K i = 600 pM); In addition, other PC's are also inhibited at higher concentrations. In biochemical, cellular and animal studies, α 1 -PDX was able to block furin activity and prevent the production of pathogenic viruses, bacterial toxin activation and cancer metastasis .

function

The protein is an enzyme that belongs to the subtilisin-like proprotein convertase family. The members of this family are proprotein convertases, which latently convert precursor proteins to their active variants. It is a calcium-dependent serine endoprotease that can very efficiently cleave precursor proteins at their paired basic amino acid processing sites. Some of the furin substrates are pro- parathyroid hormone , TGF-β1 , pro- albumin , pro- beta secretase , matrix metalloproteinase-1, beta-NGF and Von Willebrand factor . A furin-like proprotein convertase is involved in the processing of RGMc (also called hemojuvelin ), which can cause a serious condition called juvenile hemochromatosis due to iron overload. Research groups led by Ganz and Rotwein demonstrated that furin-like proprotein convertase (PPC) are responsible for the conversion of 50 kDa HJV to a 40 kDa protein with a truncated COOH terminus at a polybasic RNRR site. It suggests a potential mechanism for the generation of soluble forms of hemojuvelin (s-hemojuvelin), which can be found in the blood of rodents and humans.

Furin is one of the proteases responsible for the proteolytic cleavage of HIV - Cases polyprotein precursor gp160 to gp120 and gp41 in advance of the viral assembly. It is believed that this gene plays a role in tumor development. Uses of alternative polyadenylation sites have been found for this gene.

Furin is abundant in the Golgi apparatus, where it breaks down other proteins into their mature / active forms. Furin only cleaves proteins downstream of a basic amino acid target sequence (typically Arg-X- (Arg / Lys) -Arg '). In addition to processing cellular precursor proteins, furin is also used by several pathogens. For example, the envelope proteins of the viruses HIV, influenza , dengue fever , several filoviruses including Ebola and Marburg virus must be cleaved by furin or furin-like proteases so that they can become fully functional. In the SARS-CoV-2 virus, the involvement of furin in cell access has been demonstrated. This has a furin-affine protein expression on its spike protein and is split there by furin in order to initiate endosomal cell access in the lung tissue. Anthrax toxin , Pseudomonas - exotoxin , and papilloma viruses have to be processed by furin while the host cell to enter. Furin inhibitors are tested as therapeutic agents for the treatment of anthrax - infection .

The furin substrates and the positions of the furin cleavage sites in protein sequences can be predicted by two bioinformatic methods: ProP and PiTou.

Furin expression in T cells is necessary for the maintenance of peripheral immune tolerance .

Furin interacts with PACS1 .

Individual evidence

  1. ↑ NG Seidah, R. Day, M. Marcinkiewicz, M. Chrétien: precursor convertases: ancient in evolutionary, cell-specific, combinatorial mechanism yielding diverse bioactive peptides and proteins. In: Annals of the New York Academy of Sciences. Volume 839, May 1998, pp. 9-24, doi : 10.1111 / j.1749-6632.1998.tb10727.x , PMID 9629127 (review).
  2. ↑ C. Thacker, AM Rose: A look at the Caenorhabditis elegans Kex2 / Subtilisin-like proprotein convertase family. In: BioEssays: news and reviews in molecular, cellular and developmental biology. Volume 22, Number 6, June 2000, pp. 545–553, doi : 10.1002 / (SICI) 1521-1878 (200006) 22: 6 <545 :: AID-BIES7> 3.0.CO; 2-F , PMID 10842308 ( Review).
  3. ↑ a b G. Thomas: Furin at the cutting edge: from protein traffic to embryogenesis and disease. In: Nature reviews. Molecular cell biology. Volume 3, number 10, October 2002, pp. 753-766, doi : 10.1038 / nrm934 , PMID 12360192 , PMC 1964754 (free full text) (review).
  4. ^ RJ Siezen, JW Creemers, WJ Van de Ven: Homology modeling of the catalytic domain of human furin. A model for the eukaryotic subtilisin-like proprotein convertases. In: European Journal of Biochemistry . Volume 222, Number 2, June 1994, pp. 255-266, doi : 10.1111 / j.1432-1033.1994.tb18864.x , PMID 8020465 .
  5. ^ NC Rockwell, RS Fuller: Specific modulation of Kex2 / furin family proteases by potassium. In: Journal of Biological Chemistry . Volume 277, Number 20, May 2002, pp. 17531-17537, doi : 10.1074 / jbc.M111909200 , PMID 11893737 .
  6. ^ SS Molloy, PA Bresnahan, SH Leppla, KR Klimpel, G. Thomas: Human furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-XX-Arg and efficiently cleaves anthrax toxin protective antigen. In: Journal of Biological Chemistry . Volume 267, Number 23, August 1992, pp. 16396-16402, PMID 1644824 .
  7. ↑ JA Walker, SS Molloy, G. Thomas, T. Sakaguchi, T. Yoshida, TM Chambers, Y. Kawaoka: Sequence specificity of furin, a proprotein-processing endoprotease, for the hemagglutinin of a virulent avian influenza virus. In: Journal of Virology . Volume 68, Number 2, February 1994, pp. 1213-1218, PMID 8289354 , PMC 236564 (free full text).
  8. ↑ DJ Krysan, NC Rockwell, RS Fuller: Quantitative characterization of furin specificity. Energetics of substrate discrimination using an internally consistent set of hexapeptidyl methylcoumarinamides. In: Journal of Biological Chemistry . Volume 274, Number 33, August 1999, pp. 23229-23234, doi : 10.1074 / jbc.274.33.23229 , PMID 10438496 .
  9. ^ A b F. Jean, K. Stella, L. Thomas, G. Liu, Y. Xiang, AJ Reason, G. Thomas: alpha1-Antitrypsin Portland, a bioengineered serpin highly selective for furin: application as an antipathogenic agent. In: Proceedings of the National Academy of Sciences . Volume 95, Number 13, June 1998, pp. 7293-7298, doi : 10.1073 / pnas.95.13.7293 , PMID 9636142 , PMC 22594 (free full text).
  10. ↑ ED Anderson, L. Thomas, JS Hayflick, G. Thomas: Inhibition of HIV-1 gp160-dependent membrane fusion by a furin-directed alpha 1-antitrypsin variant. In: Journal of Biological Chemistry . Volume 268, Number 33, November 1993, pp. 24887-24891, PMID 8227051 .
  11. ^ S. Molloy, G. Thomas: The Enzymes . Academic Press, San Diego (California) 2001, pp. 199-235 .
  12. ↑ Lin L, Nemeth E, Goodnough JB, Thapa DR, Gabayan V, Ganz T: Soluble hemojuvelin is released by proprotein convertase-mediated cleavage at a conserved polybasic RNRR site . In: Blood Cells, Molecules & Diseases . 40, No. 1, 2008, pp. 122-31. doi : 10.1016 / j.bcmd.2007.06.023 . PMID 17869549 . PMC 2211380 (free full text).
  13. Jump up ↑ Kuninger D, Kuns-Hashimoto R, Nili M, Rotwein P: Pro-protein convertases control the maturation and processing of the iron-regulatory protein, RGMc / hemojuvelin . In: BMC Biochemistry . 9, 2008, p. 9. doi : 10.1186 / 1471-2091-9-9 . PMID 18384687 . PMC 2323002 (free full text).
  14. ↑ Hallenberger S, Bosch V, Angliker H, Shaw E, Klenk HD, Garten W: Inhibition of furin-mediated cleavage activation of HIV-1 glycoprotein gp160 . In: Nature . 360, No. 6402, November 1992, pp. 358-61. doi : 10.1038 / 360358a0 . PMID 1360148 .
  15. ↑ Entrez Gene: FURIN furin (paired basic amino acid cleaving enzyme) .
  16. ↑ Roebroek AJ, Schalken JA, Leunissen JA, Onnekink C, Bloemers HP, Van de Ven WJ: Evolutionary conserved close linkage of the c-fes / fps proto-oncogene and genetic sequences encoding a receptor-like protein . In: The EMBO Journal . 5, No. 9, September 1986, pp. 2197-202. PMID 3023061 . PMC 1167100 (free full text).
  17. ^ Thomas G: Furin at the cutting edge: from protein traffic to embryogenesis and disease . In: Nature Reviews Molecular Cell Biology . 3, No. 10, October 2002, pp. 753-66. doi : 10.1038 / nrm934 . PMID 12360192 . PMC 1964754 (free full text).
  18. ↑ Hoffmann, Kleine-Weber, Pöhlmann: " A Multibasic Cleavage Site in the Spike Protein of SARS-CoV-2 Is Essential for Infection of Human Lung Cells ", In: molecular-cell , May 21, 2020, doi: 10.1016 / j .molcel.2020.04.022
  19. ↑ Coutard B, Valle C, de Lamballerie X, Canard B, Seidah NG, Decroly E: The spike glycoprotein of the new coronavirus 2019-nCoV contains a furin-like cleavage site absent in CoV of the same clade . In: Antiviral Research . 176, February 2020, p. 104742. PMID 32057769 .
  20. ↑ Shiryaev SA, Remacle AG, Ratnikov BI, Nelson NA, Savinov AY, Wei G, Bottini M, Rega MF, Parent A, Desjardins R, Fugere M, Day R, Sabet M, Pellecchia M, Liddington RC, Smith JW, Mustelin T, Guiney DG, Lebl M, Strongin AY: Targeting host cell furin proprotein convertases as a therapeutic strategy against bacterial toxins and viral pathogens . In: The Journal of Biological Chemistry . 282, No. 29, July 2007, pp. 20847-53. PMID 17537721 .
  21. ↑ Duckert P, Brunak S, Blom N: Prediction of proprotein convertase cleavage sites . In: Protein Engineering, Design & Selection . 17, No. 1, January 2004, pp. 107-112. PMID 14985543 .
  22. ^ Tian S, Huajun W, Wu J: Computational prediction of furin cleavage sites by a hybrid method and understanding mechanism underlying diseases . In: Scientific Reports . February 2, 2012. doi : 10.1038 / srep00261 . PMID 22355773 . PMC 3281273 (free full text).
  23. ↑ Pesu M, Watford WT, Wei L, Xu L, Fuss I, Strober W, Andersson J, Shevach EM, Quezado M, Bouladoux N, Roebroek A, Belkaid Y, Creemers J, O'Shea JJ: T-cell-expressed Proprotein convertase furin is essential for maintenance of peripheral immune tolerance . In: Nature . 455, No. 7210, September 2008, pp. 246-50. doi : 10.1038 / nature07210 . PMID 18701887 . PMC 2758057 (free full text).
  24. ↑ Wan L, Molloy SS, Thomas L, Liu G, Xiang Y, Rybak SL, Thomas G: PACS-1 defines a novel gene family of cytosolic sorting proteins required for trans-Golgi network localization . In: Cell . 94, No. 2, July 1998, pp. 205-16. doi : 10.1016 / S0092-8674 (00) 81420-8 . PMID 9695949 .